Expert Consensus on Bilateral Inferior Petrosal Sinus Sampling in the Diagnosis of Cushing′s Syndrome (2023) / 罕见病研究

@#Cushing′s syndrome(CS)is a clinical syndrome caused by a variety of causes, with main manifestations exhibited by central obesity, purple skin striae, hypertension, and diabetes. In patients with adrenocorticotropic hormone(ACTH)-dependent CS, the result of bilateral inferior petrosal sinus sampling(BIPSS)is the gold standard for determining the source of ACTH in the absence of routine imaging findings.However, the indications, contraindications, operating procedures, precautions and outcome judgments of BIPSS differ from one medical center to another, and there are currently no international and domestic clinical guidelines and expert consensus on BIPSS. In order to further improve the operation specifications of BIPSS in the diagnosis process of difficult CS, the Hypothalamic and Pituitary Disease Group of the China Alliance for Rare Diseases and the Innovation Center of Pituitary Diseases of Peking Union Medical College Hospital organized experts in endocrinology, interventional radiology, neurosurgery and laboratory in China to formulate this consensus. This consensus aims to provide standardized methodological guidance for the use of BIPSS in the differential diagnostic process of CS in clinical practice.

Precision gene editing technologies based on CRISPR/Cas9: a review / 生物工程学报

Gene editing technology is a genetic operation technology that can modify the DNA sequence at the genomic level. The precision gene editing technology based on CRISPR/Cas9 system is a gene editing technology that is easy to operate and widely used. Unlike the traditional CRISPR/Cas9 system, the precision gene editing technology can perform site-directed mutation of genes without DNA template. This review summarizes the recent development of precision gene editing technology based on CRISPR/Cas9, and prospects the challenges and opportunities of this technology.

Multiplex gene editing and regulation techniques based on CRISPR/Cas system / 生物工程学报

The CRISPR/Cas systems comprising the clustered regularly interspaced short palindromic repeats (CRISPR) and its associated Cas protein is an acquired immune system unique to archaea or bacteria. Since its development as a gene editing tool, it has rapidly become a popular research direction in the field of synthetic biology due to its advantages of high efficiency, precision, and versatility. This technique has since revolutionized the research of many fields including life sciences, bioengineering technology, food science, and crop breeding. Currently, the single gene editing and regulation techniques based on CRISPR/Cas systems have been increasingly improved, but challenges still exist in the multiplex gene editing and regulation. This review focuses on the development and application of multiplex gene editing and regulation techniques based on the CRISPR/Cas systems, and summarizes the techniques for multiplex gene editing or regulation within a single cell or within a cell population. This includes the multiplex gene editing techniques developed based on the CRISPR/Cas systems with double-strand breaks; or with single-strand breaks; or with multiple gene regulation techniques, etc. These works have enriched the tools for the multiplex gene editing and regulation and contributed to the application of CRISPR/Cas systems in the multiple fields.

The progress of the gene editing therapy of inherited retinal diseases based on CRISPR/Cas9 / 中华眼底病杂志

Inherited retinal diseases (IRDs) are the major cause of refractory blinding eye diseases, and gene replacement therapy has already made preliminary progress in the treatment of IRDs. For IRDs that cannot be treated by gene replacement therapy, gene editing provides an alternative therapeutic method. Strategies like disruption of pathogenic variants with or without gene augmentation therapy and precise repair of pathogenic variants can be applied for IRDs with various inheritance patterns and pathogenic variants. In animal models of retinitis pigmentosa, Usher syndrome, Leber congenital amaurosis, cone rod cell dystrophy, and other disorders, CRISPR/Cas9, base editing, and prime editing showed the potential to edit pathogenic variations in vivo, indicating a promising future for gene editing therapy of IRDs.

Research progress of next-generation gene editing tools / 中国药科大学学报

@#Gene editing tools with nucleases as the main component have now implemented programmable targeted mutagenesis or insertion or deletion of mammalian genomes.From zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), CRISPR/Cas system to safer and more accurate Cas9 fusion protein gene editing tools and other nuclease gene editing tools, this paper systematically describes the development and evolution of gene editing, with detailed introduction to the development and optimization of next-generation gene editing tools, and a prospect of the clinical application of and challenges for gene editing tools.

Effects and mechanisms of SNP-9 on Aβ25-35-induced damage in bEnd.3 cells / 中国药科大学学报

@#In order to investigate the effects of neuroprotective peptide SNP-9 which is derived from silk fibroin hydrolysate on the injury of the blood-brain barrier in Alzheimer′s disease (AD), Aβ25-35 was used to damage brain microvascular endothelial cells bEnd.3 to establish AD injury model and drug intervention was performed.MTT assay was used to detect the effects of SNP-9 and Aβ25-35 on cell viability.RT-qPCR was used to determine the effects of SNP-9 and Aβ25-35 on the mRNA levels of tight junctions (TJs)-related ZO-1, occludin and claudin-5.Western blot was used to detect the effects of SNP-9 and Aβ25-35 on the protein levels of TNF-α, phosphorylated NF-κB, NF-κB, IκBα and RAGE.The results showed that SNP-9 reduced bEnd.3 cell damage induced by Aβ25-35, and improved the abnormal mRNA levels of ZO-1, occludin and claudin-5 in model cells.It alleviated the abnormal protein levels of TNF-α, phosphorylated NF-κB, IκBα and RAGE induced by Aβ25-35. These results suggest that SNP-9 may regulate the levels of TNF-α in model cells by influencing RAGE/NF-κB pathway, and then ameliorate TJs-related abnormalities and alleviate bEnd.3 cell injury induced by Aβ25-35.

Plant prime editing technique: a new genome editing tool for plants / 生物工程学报

The CRISPR/Cas9 based prime editing (PE) technique enables all 12 types of base substitutions and precise small DNA deletions or insertions without generating DNA double-strand breaks. Prime editing has been successfully applied in plants and plays important roles in plant precision breeding. Although plant prime editing (PPE) can substantially expand the scope and capabilities of precise genome editing in plants, its editing efficiency still needs to be further improved. Here, we review the development of PPE technique, and introduce structural composition, advantages and limitations of PPE. Strategies to improve the PPE editing efficiency, including the Tm-directed PBS length design, the RT template length, the dual-pegRNA strategy, the PlantPegDesigner website, and the strategies for optimizing the target proteins of PPE, were highlighted. Finally, the prospects of future development and application of PPE were discussed.

Immunogenicity of chimeric peptide gene of Xinjiang hemorrhagic fever virus glycoprotein / 国际生物医学工程杂志

Objective:To investigate the effects of different chimerism strategies and different immune ways on the two antigen-dominant regions of Xinjiang hemorrhagic fever virus (XHFV) glycoprotein.Methods:The 5' end was added or not added with interleukin-2 (IL-2) signal peptide and the general-purpose auxiliary T cell epitopes as different design strategies. GcⅠ and GcⅡ and the epitopes previously identified on GcⅠ (Gc 233-248, Gc 241-256 and Gc 281-296) were fused and constructed into the eukaryotic expression vector pVAX1 and the prokaryotic expression vector pET-28a. The recombinant prokaryotic plasmid transformed into E.coli BL21 was induced and purified, and the recombinant eukaryotes were extracted by indirect immunofluorescent assay. BALB/c mice were immunized by protein immunity, gene immunity, and DNA prime-protein boost immunity. The IgG antibody level was measured by ELISA. The immune effect was evaluated by the proliferation of T-lymphocytes and the content of cytokines in the spleen. Results:The results of double enzyme digestion and sequencing showed that eight recombinant plasmids were successfully constructed, and the recombinant eukaryotes were successfully expressed in vitro by fluorescence microscopy. After three times of immunization, the IgG level and the proliferation of T-lymphocytes in the spleen of mice in the experimental group were significantly higher than those in the control group ( P<0.01). The mass concentration test results of Th2 cytokines IL-4 and Th1 cytokines interferon-gamma (IFN-γ) revealed that the response of the DNA prime-protein boost immunity was biased to Th1. Conclusions:The multi-epitope chimeric vaccine of XHFV glycoprotein was successfully constructed, and the target antigen could be expressed effectively in vivo. The immune groups stimulated stronger humoral and cellular immune responses compared with the control group. Among them, the immune effect of pVAX1-ST(GcⅠe+GcⅡ) combined with recombinant protein r(GcⅠe+GcⅡ) was the best, and it is expected to be a new candidate vaccine for XHFV.

Role of hepatic H19 expression in glucose metabolism disorder inducedby p,p'-dichlorodiphenyldichloroethylene / 预防医学

Objective @#To investigate the role of hepatic long-chain non-coding RNA (lncRNA) H19 in key genes associated with glucose metabolism disorder induced by p,p′-dichlorodiphenyldichloroethylene (p,p′-DDE).@*Methods@#Human embryonic liver CCC-HEL-1 cells were divided into the DMSO group, 0.1 μmol/L p,p′-DDE group, 1 μmol/L p,p′-DDE group, 10 μmol/L p,p′-DDE group, small interference RNA (siRNA)+DMSO group and siRNA+10 μmol/L p,p′-DDE group. The promoter region methylation, mRNA expression and protein expression of hepatocyte nuclear factor 4α (HNF4α), forkhead box transcription factor O1 (FoxO1) and insulin-like growth factor (IGF2) were detected in CCC-HEL-1 cells using the bisulfite method, real-time fluorescence quantitative PCR (qPCR) assay and BCA assay, respectively. The changes in H19 mRNA expression, the methylation of associated genes in the promoter region and transcriptional expression were compared in CCC-HEL-1 among groups.@*Results@#Exposure to p,p′-DDE alone at different doses resulted in an increase in H19 expression, and the H19 mRNA expression was higher in the 10 μmol/L p,p′-DDE group than in the DMSO group [(1.31±0.25) vs. (1.02±0.22); P<0.05]. Lower methylation of the HNF4α gene in the pro moter region [(38.59±32.77)% vs. (61.43±24.64)%; P<0.05] and higher HNF4α mRNA expression [(1.33±0.26) vs. (1.03±0.28); P<0.05] were detected in the 10 μmol/L p,p′-DDE group than in the DMSO group, while no significant differences were detected between the two groups in terms of the methylation of FoxO1 and IGF2 genes in the promoter region, FoxO1 and IGF2 mRNA and protein expression (P>0.05). Following siRNA-induced H19 knockdown, higher methylation of the HNF4α gene in the promoter region [(71.33±22.23)% vs. (38.59±32.78)%; P<0.05], lower HNF4α mRNA expression [(0.71±0.17) vs. (1.33±0.26); P<0.05], higher methylation of FoxO1 gene in the promoter region [(47.73±34.24)% vs. (25.09±25.35)%; P<0.05] and higher IGF2 mRNA [(1.39±0.25) vs. (0.80±0.20); P<0.05] were found in the siRNA+DMSO group than in the 10 μmol/L p,p′-DDE group, and higher IGF2 protein expression was detected in the siRNA+DMSO group than in the DMSO group [(1.03±0.11) vs. (0.74±0.12); P<0.05]. @*Conclusion@#Hepatic H19 may alter HNF4α, FoxO1 and IGF2 transcription and expression through mediating the methylation of genes in the promoter region, thereby playing a role in p,p′-DDE-induced glucose metabolism disorders.

A Role of TREX1 in Immune Regulation and Human Diseases / 中国生物化学与分子生物学报

Three prime repair exonuclease 1 (TREX1), also known as DNase IE, is a major 3'-5' restriction exonuclease in most of tissues and cell types of the mammals. The exonuclease activity of TREX1 plays an essential role in maintaining the immune tolerance of the innate immune system, which avoids the excessive activation of the innate immune system and massive production of auto-antibodies induced by the abnormal accumulation of cytosolic DNA. cGAS-STING signaling was identified as an important innate immune response to pathogens and maintained cellular environmental homeostasis. TREX1 prevents occasional leakage of nuclear DNA into the cytosol, which activates cGAS and triggers the downstream type I interferons cascade. Mutations of human TREX1 cause a series of autoimmune diseases, such as Aicardi-Goutieres syndrome (AGS), Familial chilblain lupus (FCL), Systemic lupus erythematosus (SLE) and Leukodystrophy-related retinopathy (RVCL). Besides, TREX1 inhibits the innate immune response to human immunodeficiency virus type 1 (HIV-1) and plays an important role in mediating the viral immune evasion. Moreover, TREX1 acts as an upstream regulator of the DNA sensing pathway, which maintains tumor immune tolerance and prevents cell senescence. Here, we focus on the immune regulation of TREX1 and demonstrate the role of TREX1 in autoimmune diseases, HIV-1 infection, cancer and cell senescence to provide the basic theoretical guidance for human disease therapy.

Effects and mechanisms of microRNA-23b on neuronal apoptosis induced by Aβ25-35 / 中国药科大学学报

@#In this study, the effects and mechanisms of miR-23b in the AD cell model were explored. Aβ25-35 was used to induce neuronal injury model, and cell viabilities were detected by MTT assay. The effect of miR-23b on the apoptotic levels of Aβ25-35-induced SH-SY5Y cells was analyzed using Annexin V-FITC/PI detection kit. The effect of miR-23b on the mitochondrial membrane potential of Aβ25-35-induced SH-SY5Y cells was examined using JC-1 fluorescent probe. The levels of cell apoptosis-related proteins and autophagy-related proteins were detected by Western blot. The results showed that miR-23b could alleviate the apoptosis and the abnormal mitochondrial membrane potential in SH-SY5Y cells induced by Aβ25-35.This study suggested that miR-23b may attenuate Aβ25-35-induced neuronal apoptosis by regulating apoptosis and autophagy-related pathway.

Evaluation of the efficacy of cevimeline in improving the clinical symptoms of patients with primary Sjögren′s syndrome / 口腔疾病防治

Objective@# To investigate the clinical efficacy of cevimeline as a pharmacotherapeutic approach to stimulating gland activity in improving the symptoms and signs of primary Sjögren syndrome (pSS).@*Methods@#Sixty-three patients diagnosed with pSS who attended the Affiliated Huai'an Hospital of Xuzhou Medical University from January 2018 to September 2019 were included in this trial. They were randomly assigned to the therapeutic group and control group. All patients were recalled at baseline and after 2 weeks, 3 months and 6 months. Measurement of salivary and lacrimal flow as well as evaluation of subjective symptoms was performed at the follow-up. @*Results@# Fifty-eight patients completed the trial and were included in the statistical analysis. There was a significant difference between the two groups in the measurement of salivary and lacrimal flow at the second week and third month (P < 0.05). Improvement in subjective symptoms of oral, ocular and gland was detected at the third month (P < 0.05). At the sixth month, compared with the control group, only the salivary gland symptom score of the treatment group was statistically significant (P < 0.05). @*Conclusion@#Cevimeline has good specificity and safety and can increase salivary and lacrimal flow and improve subjective symptoms of pSS in a short time.

Research progress on the role of T helper cells 17 and interleukin 17 in oral mucosal diseases / 口腔疾病防治

@#T helper 17 (Th17) cells are a new type of CD4+ T helper cell. They participate in the immune and inflammatory response by secreting specific interleukin-17 (IL-17). In oral mucosal diseases, oral lichen planus (OLP), recurrent aphthous ulcer (RAU) and Behcet′s disease (BD) are associated with Th17 cells and IL-17. There were 17 kinds of proteins in the saliva of patients with OLP that could upregulate the expression of Th17 cells and induce the secretion of IL-17. IL-17 can stimulate epithelial cells, endothelial cells and fibroblasts to produce a variety of cytokines, such as IL-6, IL-8, granulocyte macrophage colony-stimulating factor and cell adhesion molecule-1, leading to the production and aggravation of inflammation. Th17/Tc17 cell-targeted therapy can significantly improve the clinical symptoms of OLP patients′ mucosa and skin. IL-17 can stimulate oral keratinocytes through the IL-17RA or IL-17RE receptor and produce proinflammatory effects in RAU. Th17 cells in the peripheral blood of BD patients are significantly increased, while Treg cells are significantly decreased.

CRISPR/Cas-mediated DNA base editing technology and its application in biomedicine and agriculture / 生物工程学报

In recent years, the genome editing technologies based on the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) have developed rapidly. The system can use homologous directed recombination (HDR) to achieve precise editing that it medicated, but the efficiency is extremely low, which limits its application in agriculture and biomedical fields. As an emerging genome editing technology, the CRISPR/Cas-mediated DNA base editing technologies can achieve targeted mutations of bases without generating double-strand breaks, and has higher editing efficiency and specificity compared with CRISPR/Cas-mediated HDR editing. At present, cytidine base editors (CBEs) that can mutate C to T, adenine base editors (ABEs) that can mutate A to G, and prime editors (PEs) that enable arbitrary base conversion and precise insertion and deletion of small fragments, have been developed. In addition, glycosylase base editors (GBEs) capable of transitioning from C to G and double base editors capable of editing both A and C simultaneously, have been developed. This review summarizes the development, advances, advantages and limitations of several DNA base editors. The successful applications of DNA base editing technology in biomedicine and agriculture, together with the prospects for further optimization and selection of DNA base editors, are discussed.

Therapeutic effect and mechanism of sodium formononetin-3′-sulphonate on collagen-induced rheumatoid arthritis / 中国药科大学学报

@#To investigate the therapeutic effect and mechanism of sodium formononetin-3′-sulphonate (SFS) on collagen-induced rheumatoid arthritis in mice, C57 mice were induced with chicken type II collagen to establish a model of rheumatoid arthritis (collagen-induced arthritis, CIA), and were injected intraperitoneally with different doses of SFS (50,100,200 mg/kg). Body weight, food intake and foot swelling of all groups were observed during the experiment.After the treatment, TNF-α, IL-6, and IL-10 in the serum were detected with the CBA kit; NF-κB p65, p-NF-κB p65 (p-p65), TIPE2, PCNP and IκB-α in spleen tissue were determined by Western blot; the organ index, pathological changes of ankle joint cartilage tissue and the positive expression of NF-κB p65 in ankle joint tissue were also observed.The results showed that, compared with the model group, the body weight and food intake of mice in the treatment group increased, while the degree of foot swelling decreased; the expression levels of inflammatory factors TNF-α and IL-6 in serum decreased, while the expression of anti-inflammatory factor IL-10 increased; the expression levels of NF-κB p65, p-p65 and PCNP in spleen tissue decreased, while the expression of TIPE2 and IκB-α protein increased; the index of spleen and thymus of the CIA mice in the treatment group, the infiltration of inflammatory cells in the ankle joint, the destruction of synovial tissue and cartilage, and the positive expression of NF-κB p65 decreased.Among them, the high-dose group of SFS showed a better therapeutic effect.It is suggested that SFS has a therapeutic effect on CIA mice, and the mechanism may be achieved by regulating the NF-κB p65 signaling pathway and inhibiting the expression of inflammatory factors.

Assessment of the acid phosphatase CP01850 from Corynebacterium pseudotuberculosis in DNA and subunit vaccine formulations against caseous lymphadenitis / Avaliação da fosfatase ácida recombinante CP01850 de Corynebacterium pseudotuberculosis em formulações vacinais de subunidade e de DNA contra a linfadenite caseosa

The target cp1002_RS01850 from Corynebacterium pseudotuberculosis was used to construct a DNA and recombinant subunit vaccine against caseous lymphadenitis. Recombinant protein rCP01850 was expressed in Escherichia coli using pAE vector, and DNA vaccine was engineered with pTARGET vector. BALB/c mice were divided in five groups containing eight animals each, inoculated with: pTARGET/cp01850 as DNA vaccine (G1); rCP01850 plus Al (OH)3 as recombinant subunit vaccine (G2); pTARGET/cp01850 and a boost with rCP01850 plus Al (OH)3 (G3); pTARGET (G4); or Al (OH)3 (G5). Mice were inoculated and blood samples were collected on days 0, 21, and 42 for the analysis of total IgG, IgG1 and IgG2a by ELISA. In each group, five animals were challenged with Mic-6 C. pseudotuberculosis strain, and three were used for cytokine quantification by qPCR. Although no group has been protected by vaccines against lethal challenge, G2 showed an increase in the survival rate after challenge. Significantly higher levels of IL-4, IL-12, IFN-γ, total IgG, IgG1 and IgG2a were also detected for G2, evidencing a mixed Th1/Th2 immunological profile. In conclusion, despite no protection level provided by different vaccinal strategies using cp1002_RS01850 from C. pseudotuberculosis, G2 developed a Th1/Th2 immune response with an increase in survival rate.(AU)

O alvo cp1002_RS01850 de Corynebacterium pseudotuberculosis foi utilizado para construir uma vacina recombinante de subunidade e de DNA contra a linfadenite caseosa. A proteína recombinante rCP01850 foi expressa em Escherichia coli usando o vetor pAE, e a vacina de DNA foi construída com o vetor pTARGET. Camundongos BALB/c foram divididos em grupos de oito animais, inoculados com: pTARGET/cp01850 como vacina de DNA (G1); rCP01850 e Al (OH)3 como vacina recombinante de subunidade (G2); pTARGET/cp01850 e um boost com rCP01850 e Al (OH)3 (G3); pTARGET (G4); ou Al (OH)3 (G5). Os animais foram inoculados e amostras de sangue foram coletadas nos dias 0, 21, e 42 do experimento para a análise de IgG total, IgG1 e IgG2a por ELISA. De cada grupo, cinco animais foram desafiados com a cepa Mic-6 de C. pseudotuberculosis, e três foram usados para a quantificação de citocinas por qPCR. Apesar de nenhum grupo ter sido protegido pelas vacinas testadas contra o desafio letal, G2 apresentou taxa de sobrevida e níveis de IL-4, IL-12, IFN-γ, IgG total, IgG1 e IgG2a significativamente mais altos, evidenciando um perfil imunológico misto Th1/Th2. Conclui-se que apesar das diferentes estratégias vacinais utilizando cp1002_RS01850 de C. pseudotuberculosis não terem sido capazes de gerar proteção, G2 desenvolveu uma resposta Th1/Th2 e elevou a taxa de sobrevida.(AU)

Influence of two kinds of zirconia primers on the bonding effect of zirconia ceramics / 口腔疾病防治

Objective@#To evaluate the effect of two commonly used zirconia primers on the bond strength and durability of zirconia ceramics. @*Methods@# Zirconia wafers with a diameter of 5 mm and a thickness of 2 mm were cut and prepared by CAD/CAM (51). After sintering, sandblasting, ultrasonic cleaning and drying, the zirconia wafers are divided into three groups according to the types of zirconia primer: group A, no primer group; group B, Z-Prime plus; and group C, Clearfil Ceramic Primer, with each set containing 17 samples. The surface morphology of zirconia was observed by scanning electron microscopy after applying the coating primer according to the operation specifications. Then, the microshear bonding specimens were made, and the bonding strength was tested after water storage for 3 days and 5 000 × thermocycling. @*Results@#The surface of zirconia ceramics was roughened by sandblasting. After Z-Prime plus coating, the primer did not completely cover the zirconia surface, which is island like. After coating with the Clearfil Ceramic Primer, the primer was completely covered on the surface of zirconia ceramics without exposure to zirconia. Energy spectrum analysis showed that the content of C, O, Si and P in the coating area was relatively high. After primer coating, the bonding strength between zirconia and resin cement was significantly higher than that of group A (20.6 ± 2.1)MPa, P < 0.05, while there was no significant difference in the immediate bonding strength between group B (33.2 ± 3.9)MPa and group C (30.7 ± 2.4)MPa (P > 0.05). After 5 000 × thermocycling, the bonding strength of group A was(4.1 ± 2.5)MPa, that of group B was (23.1 ± 2.3)MPa and that of group C was (28.9 ± 2.6)MPa. There were statistically significant differences among groups A, B and C. The aging adhesive strength of groups A and B was significantly reduced compared with the immediate adhesive strength (P < 0.05). Group C had the highest bonding strength after aging (28.9 ± 2.6) MPa, with no significant difference before and after aging (P > 0.05). After 5 000 × thermocycling aging, the percentage of bonding failure in the fracture mode of group A increased from 66% to 100% and that of group B increased from 16% to 53%. In group C, the percentage of bonding failure increased from 20% to 24%.@*Conclusion @# The zirconia primers on the surface of zirconia ceramics after sandblasting can enhance the bonding strength and durability of zirconia.

Advances of near-infrared fluorescent probes for detection of Alzheimer′s disease / 中国药科大学学报

@#Alzheimer′s disease(AD)is a chronic neurodegenerative diseasecommonly seen in the elderlys. Several therapeutic drugs have failed in phase III clinical trials in recent years and there have been no efficient treatment for AD currently. Thus, there has been an urgent need for the effective methods of AD diagnosis at early stage. Developingnear-infrared fluorescentprobes for AD hallmarks detection has been a promising research field. In this review, we summarized a variety of near-infrared fluorescence(NIRF)probes reported in the past decade, which capable of detecting β-amyloid, Tau protein and reactive oxygen species, including their chemical strucutres、optical properties, in vitro and in vivo studies. Furthermore, we alsoraised some new directions for AD diagnosing. We believe that these new directions raised herein will benefit the future development of NIRF probes in the field of AD research.

Whole Genome Bisulfite Sequencing Study on Parkinson′s Disease Model Mice Genetically Modified by A53T Mutant α-Synuclein in Midbrain / 中山大学学报(医学科学版)

@#【Objective】To investigate the effect of PD-related gene SNCA mutated in A53T on methylation modification in the dopaminergic neurons from the mouse midbrain.【Methods】The midbrain tissue from the A53T mutant human α-synuclein（hA53T α-syn）transgenic mice and non-transgenic（nTg）mice were isolated α-synmice. Bisulfite-sequencing（BS-seq）was utilized for analyzing the DNA methylation of 12-month-old of hA53T α-synmice and nTg mice at a whole genome level. Subsequently，differentially methylated regions（DMRs）were screened for GO enrichment analyses.【Results】Through comparative analyses，481 DMRs were found. Among the data ，hypermethylated and hypomethylated DMRs accounted for 257 and 224 respectively. These DMRs involved in ubiquitin degradation pathway-related genes， including Ubqln2，HECTD4，Rnf157 genes；serine/threonine protein kinase PINK1 gene，etc. Enrichment data revealed that the genes containing DMRs projected to 545 GO sub-terms，and significantly enriched in anatomical structure development，dendrite development，nervous system development，neuronal projection，etc.【Conclusion】The A53T mutation of SNCA gene which is related to PD could introduce DNA methylation alterations in mouse midbrain.

Research status of IgG4⁃related sialadenitis / 口腔疾病防治

@#IgG4⁃related sialadenitis (IgG4⁃RS) is a type of autoimmune disease that has been recognized in recent years, and the pathogenesis remains unclear. IgG4⁃RS mainly affects the submandibular gland and parotid gland and is characterized by diffuse painless swelling of the bilateral salivary glands and/or lacrimal glands, usually lasting more than 3 months. Some patients have accompanying hearing loss or hearing impairment, sinusitis, lymphadenopathy and other symptoms; nearly half of patients have different degrees of salivary gland secretion disorders. Most patients have elevated serum IgG4 levels, but they cannot be used as the only marker for diagnosis. Histopathology remains the“gold standard”for diagnosis. Presently, submandibular gland biopsy is often used for diagnosis. Histopathology showed lym⁃phoplasmacytic infiltration, occlusive phlebitis, striated fibrosis; immunohistochemistry showed IgG4 + /IgG + plasma cells >40%, and IgG4 + plasma cell/high⁃power field vision > 10. Glucocorticoids are regarded as first⁃line drugs for the treat⁃ment of this disease. Clinically, glucocorticoids are often combined with immunosuppressive agents such as cyclophos⁃phamide, but no standard drug regimen exists. Most patients have a significant short⁃term treatment effect, and the long⁃term prognosis requires further study. Patients with a recurrence tendency should adjust the hormone dose over time. In the future, further research is needed regarding the pathogenesis and treatment of the disease to improve the clinical di⁃agnosis rate and therapeutic effect.